cDNA cloning and expression of a Talaromyces emersonii β-glucosidase determinant in Escherichia coli

Morrison, Jacinta; Jackson, Elizabeth A.; Bunni, L.; Coleman, David C.; McHale, Anthony P.

doi:10.1016/0167-4781(90)90080-l

Cited by 6 publications

(3 citation statements)

References 16 publications

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“…A number of R. emersonii cellulosedegrading enzymes have previously been characterized (Coughlan et al, 1984;Moloney et al, 1983;McHale, 1987), including four endoglucanases and three cellobiohydrolases. In addition, four R. emersonii BGLs have been identified and expressed (McHale & Coughlan, 1981, 1982Coughlan & McHale, 1988), of which three are GH3 enzymes: a -xylosidase (Bxl1), an avenacinase (Aven1; Morrison et al, 1990;Reen et al, 2003;Murray et al, 2004;Collins et al, 2007) and a BGL (Cel3A), which is the subject of this study. The latter has been biochemically characterized in a study by Murray et al (2004).…”

Section: Improvement Of Cellulase Mixtures: Ratio Optimization Protementioning

confidence: 99%

Structural and functional studies of the glycoside hydrolase family 3 β-glucosidase Cel3A from the moderately thermophilic fungusRasamsonia emersonii

Gudmundsson

Hansson

Karkehabadi

et al. 2016

Acta Cryst Sect D Struct Biol

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The filamentous fungus Hypocrea jecorina produces a number of cellulases and hemicellulases that act in a concerted fashion on biomass and degrade it into monomeric or oligomeric sugars. -Glucosidases are involved in the last step of the degradation of cellulosic biomass and hydrolyse the -glycosidic linkage between two adjacent molecules in dimers and oligomers of glucose. In this study, it is shown that substituting the -glucosidase from H. jecorina (HjCel3A) with the -glucosidase Cel3A from the thermophilic fungus Rasamsonia emersonii (ReCel3A) in enzyme mixtures results in increased efficiency in the saccharification of lignocellulosic materials. Biochemical characterization of ReCel3A, heterologously produced in H. jecorina, reveals a preference for disaccharide substrates over longer gluco-oligosaccharides. Crystallographic studies of ReCel3A revealed a highly N-glycosylated three-domain dimeric protein, as has been observed previously for glycoside hydrolase family 3 -glucosidases. The increased thermal stability and saccharification yield and the superior biochemical characteristics of ReCel3A compared with HjCel3A and mixtures containing HjCel3A make ReCel3A an excellent candidate for addition to enzyme mixtures designed to operate at higher temperatures.

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Section: Improvement Of Cellulase Mixtures: Ratio Optimization Protementioning

confidence: 99%

Structural and functional studies of the glycoside hydrolase family 3 β-glucosidase Cel3A from the moderately thermophilic fungusRasamsonia emersonii

Gudmundsson

Hansson

Karkehabadi

et al. 2016

Acta Cryst Sect D Struct Biol

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“…2000), only a few researches on molecular cloning have been reported from thermophilic fungi, such as Thermoascus aurantiacus (Hong et al. 2003a,b), Talaromyces emersonii (Morrison et al. 1990; Murray et al.…”

Section: Introductionmentioning

confidence: 99%

“…In recent decades, there has been increasing interest in cellulases from thermophilic fungi, which were expected to produce thermostable enzymes. Although most work has focused on purification and characterization of cellulases from thermophilic fungi (Canevascini 1999;Maheshwari et al 2000), only a few researches on molecular cloning have been reported from thermophilic fungi, such as Thermoascus aurantiacus (Hong et al 2003a,b), Talaromyces emersonii (Morrison et al 1990;Murray et al 2003Murray et al , 2004Grassick et al 2004), Humicola insolens (Varrot et al 1999;Xu et al 2001;Moriya et al 2003), Melanocarpus albomyces (Haakana et al 2004) and Humicola grisea var. thermoidea (Azevedo et al 1990;Takashima et al 1996;Pocas-Fonseca et al 1997Paula et al 2003).…”

Section: Introductionmentioning

confidence: 99%

Cloning of a gene encoding thermostable cellobiohydrolase from the thermophilic fungusChaetomium thermophilumand its expression inPichia pastoris

et al. 2009

Journal of Applied Microbiology

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Aims: A new cellobiohydrolase (CBH) gene (cbh3) from Chaetomium thermophilum was cloned, sequenced and expressed in Pichia pastoris. Methods and Results: Using RACE‐PCR, a new thermostable CBH gene (cbh3) was cloned from C. thermophilum. The cDNA of the CBH was 1607 bp and contained a 1356 bp open reading frame encoding a protein CBH precursor of 451 amino acid residues. The mature protein structure of C. thermophilum CBH3 only comprises a catalytic domain and lacks cellulose‐binding domain and a hinge region. The gene was expressed in P. pastoris. The recombinant CBH purified was a glycoprotein with a size of about 48·0 kDa, and exhibited optimum catalytic activity at pH 5·0 and 60 °C. The enzyme was more resistant to high temperature. The CBH could hydrolyse microcrystalline cellulose and filter paper. Conclusions: A new thermostable CBH gene of C. thermophilum was cloned, sequenced and overexpressed in P. pastoris. Significance and Impact of the Study: This CBH offers an interesting potential in saccharification steps in both cellulose enzymatic conversion and alcohol production.

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Expression and characterization of a cellobiohydrolase (CBH7B) from the thermophilic fungus Thielavia terrestris in Pichia pastoris

Woon

Mackeen

Mahadi

et al. 2015

Biotech and App Biochem

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The gene encoding a cellobiohydrolase 7B (CBH7B) of the thermophilic fungus Thielavia terrestris was identified, subcloned, and expressed in Pichia pastoris. CBH7B encoded 455 amino acid residues with a molecular mass of 51.8 kDa. Domain analysis indicated that CBH7B contains a family 7 glycosyl hydrolase catalytic core but lacks a carbohydrate-binding module. Purified CBH7B exhibited optimum catalytic activity at pH 5.0 and 55 °C with 4-methylumbelliferryl-cellobioside as the substrate and retained 85% of its activity following 24 H incubation at 50 °C. Despite the lack of activity toward microcrystalline substrates, this enzyme worked synergistically with the commercial enzyme cocktail Cellic CTec2 to enhance saccharification by 39% when added to a reaction mixture containing 0.25% alkaline pretreated oil palm empty fruit bunch (OPEFB). Attenuated total reflectance Fourier transform infrared spectroscopy suggested a reduction of lignin and crystalline cellulose in OPEFB samples supplemented with CBH7B. Scanning electron microscopy revealed greater destruction extent of OPEFB strands in samples supplemented with CBH7B as compared with the nonsupplemented control. Therefore, CBH7B has the potential to complement commercial enzymes in hydrolyzing lignocellulosic biomass.

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cDNA cloning and expression of a Talaromyces emersonii β-glucosidase determinant in Escherichia coli

Cited by 6 publications

References 16 publications

Structural and functional studies of the glycoside hydrolase family 3 β-glucosidase Cel3A from the moderately thermophilic fungusRasamsonia emersonii

Structural and functional studies of the glycoside hydrolase family 3 β-glucosidase Cel3A from the moderately thermophilic fungusRasamsonia emersonii

Cloning of a gene encoding thermostable cellobiohydrolase from the thermophilic fungusChaetomium thermophilumand its expression inPichia pastoris

Expression and characterization of a cellobiohydrolase (CBH7B) from the thermophilic fungus Thielavia terrestris in Pichia pastoris

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